Fixation of Brain and Incisor Teeth for Light Microscopy; Glutaraldehyde Perfusion Followed by Immersion in Bouin's Fluid

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Abstract
Fixation of brains and incisor teeth was carried out in two steps. Rats were perfused with 300 ml, mice with 100 ml, of a slightly hypertonic (390 mosM) phosphate-buffered 2-5% glutaraldehyde solution, pH 7.1. The tissues were then removed and immersed 24-48 hr in Bourn's fluid for additional fixation. Routine embedding in paraffin was used for the brain. The teeth were decalcified in disodium EDTA and dehydrated in alcohol for double embedding in celloidin-paraffin. Both tissues could be readily cut, mounted and stained by a variety of methods. Examination by light microscopy showed better architectural preservation of the tissues and more precise cytological detail than obtained by routine immersion fixation.

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